US6750031B1 - Displacement assay on a porous membrane - Google Patents

Displacement assay on a porous membrane Download PDF

Info

Publication number
US6750031B1
US6750031B1 US08/583,912 US58391296A US6750031B1 US 6750031 B1 US6750031 B1 US 6750031B1 US 58391296 A US58391296 A US 58391296A US 6750031 B1 US6750031 B1 US 6750031B1
Authority
US
United States
Prior art keywords
membrane
sample
analyte
target analyte
labelled
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Expired - Fee Related, expires
Application number
US08/583,912
Inventor
Frances S. Ligler
Anne W. Kusterbeck
Sina Y. Rabbany
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
US Secretary of Navy
Original Assignee
US Secretary of Navy
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by US Secretary of Navy filed Critical US Secretary of Navy
Priority to US08/583,912 priority Critical patent/US6750031B1/en
Assigned to NAVY, THE UNITED STATES OF AMERICA AS REPRESENTED BY THE SECRETARY OF THE reassignment NAVY, THE UNITED STATES OF AMERICA AS REPRESENTED BY THE SECRETARY OF THE ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: RABBANY, SINA Y., KUSTERBECK, ANNE W., LIGLER, FRANCES L.
Priority claimed from CA 2241324 external-priority patent/CA2241324C/en
Application granted granted Critical
Publication of US6750031B1 publication Critical patent/US6750031B1/en
Adjusted expiration legal-status Critical
Application status is Expired - Fee Related legal-status Critical

Links

Images

Classifications

    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366Apparatus specially adapted for solid-phase testing
    • G01N33/54386Analytical elements

Abstract

Displacement assays, under non-equilibrium conditions, are performed by flowing a liquid sample through a membrane having binding elements with binding sites saturated with a labelled form of the analyte. Analyte in the sample displaces, under non-equilibrium conditions, the labelled form of the analyte from the membrane. The displaced labelled form of the analyte may then be detected.

Description

BACKGROUND OF THE INVENTION

1. Field of the Invention

The present invention relates generally to assays and more specifically to displacement-type assays.

2. Description of the Background Art

U.S. Pat. No. 5,183,740, incorporated in its entirety herein for all purposes, describes a flow immunoassay system and method for performing displacement immunoassays. In a displacement assay, unlike a competitive assay, the antibody is exposed to labelled analyte prior to exposure to analyte. The analyte is in contact with the antibody and labelled, bound analyte an insufficient amount of time to establish equilibrium.

Because no time needs to be dedicated to establishing equilibrium, displacement assays are faster than competitive assays. A displacement assay, however, generally provides a smaller signal than a competitive assay. In a displacement assay, the available binding sites of the antibody are saturated or nearly saturated with labelled analyte before the unlabelled analyte is added. Since equilibrium (with labelled analyte and unlabelled analyte continually binding, releasing and competing with each other for rebinding to the available binding sites on the antibody in a steady state) has not been achieved, most of the labelled analyte in a displacement assay remains bound to the antibody and unable to provide a signal.

The relatively small signal provided by the displacement assay places an additional value on assuring the consistency of assay conditions. The bead-containing columns described in U.S. Pat. No. 5,183,740 for displacement assays must be carefully stored, prepared, and loaded to assure chemical and physical consistency (i.e., porosity, avoidance of channeling) from test to test. The need for this careful preparation and testing increases the labor, skill, and costs needed to perform accurate displacement assays. Additionally, the problems associated with the use of bead-containing columns limit the lower detection limit for displacement assays.

In studies performed at US Drug Testing, Inc. (Rancho Cucamonga, Calif.), better results for a displacement assay were achieved using tall, thin columns of beads coated with an antibody and labelled antigen than with short, wide columns. Furthermore, the efficiency with which the labelled antigen dissociated from antibody in the presence of unlabelled antigen was greater when flow rates were reduced and the antigen had more time to interact with the immobilized complex (Wemhoff et al. J. Immunol. Methods, 223-230, 1992). Both of these sets of experiments suggested that immobilization of the antibody and labelled antigen on a porous membrane would not provide a suitable matrix for the displacement assay since this geometry would not allow sufficient time, under flow conditions, for the antigen to interact efficiently with the complex to displace detectable amounts of the labelled antigen.

U.S. Pat. No. 5,369,007, to David A. Kidwell discloses a displacement assay in which samples pass through a membrane having an antibody immobilized thereon. The binding sites of the immobilized antibody are bound to an enzymatically labelled analyte. Analyte from the sample displaces the labelled analyte, causing the labelled analyte and the remainder of the sample to pass into a superabsorbent layer. The superabsorbent layer contains a substrate for the enzymatic label and any needed indicator. The Kidwell patent, however, teaches the need for a flow rate of about 0.02 ml/min and interaction times of about 1 to 5 min to assure a detectable interaction between the analyte and the antibody. In many situations, even faster results are desirable. Additionally, the Kidwell microassay card is not reusable.

SUMMARY OF THE INVENTION

Accordingly, it is an object of this invention to perform bioassays capable of detecting minute quantities of an analyte in under one minute.

It is another object of the present invention to quickly perform bioassays in a format that allows reuse of the matrix that selectively binds the analyte.

These and additional objects of the invention are accomplished by quickly flowing a sample past a non-absorbent membrane having a binding element covalently bound thereto to form attachment sites for the analyte. The available attachment sites are essentially saturated with a labelled form of the analyte. Nonspecific binding sites are blocked to prevent nonspecific binding. Additionally, the sample flows past the membrane at a rate greater than that needed to achieve equilibrium between the dissociation of labelled analyte from the binding sites and the attachment of analyte (labelled or unlabelled) thereto. The processed sample is then analyzed for the presence of any labelled antigen that the unlabelled analyte has displaced from its binding site. This analysis can be qualitative or quantitative.

BRIEF DESCRIPTION OF THE DRAWINGS

A more complete appreciation of the invention will be readily obtained by reference to the following Description of the Preferred Embodiments and the accompanying drawings in which like numerals in different figures represent the same structures or elements, wherein:

FIG. 1 schematically illustrates a device according to the present invention.

FIG. 2 schematically illustrates an alternative embodiment of a device according to the present invention.

FIG. 3 schematically illustrates another alternative embodiment of a device according to the present invention.

FIG. 4 is a graph of data from a membrane assay, in accordance with the present invention, in which the membrane was prepared by the test tube incubation method.

FIG. 5 is a graph of data from a membrane assay, in accordance with the present invention, in which the membrane was prepared by saturating the immobilized antibody with labelled analyte in the column as opposed to in a test tube.

FIG. 6 is a graph of data from a single membrane assay, according to the present invention, prepared by saturating the antibody directly in the column.

FIG. 7 is a flowchart schematically illustrating an embodiment of an assay according to the method of the present invention.

FIGS. 8a, 8 b, and 8 c show the results obtained from assay performed in accordance with the method flowcharted in FIG. 7.

DESCRIPTION OF THE PREFERRED EMBODIMENTS

Membranes useful in the present invention are typically non-absorbent (with respect to aqueous materials) materials. The non-absorbent membrane assists in providing a fast flow-through rate. Additionally, the use of a non-absorbent membrane allows the membrane, once used, to be readily rinsed of sample and reused. If displacement has occurred, reloading with labelled analyte is an option.

Typically, membranes useful in the present invention have thicknesses, exposed surface areas, and porosities that allow detection of the analyte with an interaction time of about 0.1 sec to about 30 seconds, and typically about 1 sec to about 15 seconds, between a sample suspected of containing of the analyte and the membrane having a labelled analyte of the analyte thereon. Generally, the pore sizes in the membrane are about 0.2-1.0 microns, and are typically about 0.45 microns. Of course, other pore sizes may be used to achieve the desired interaction time. Likewise, the thickness and surface area of the membrane can be adjusted to provide the desired interaction time.

Any non-absorbent membrane, of appropriate pore size and density of sites for immobilizing binding elements for the analyte, may be used. For example, the membrane may be a polyamide (e.g., Nylon™ membranes such as Immunodyne ABC™(a Nylon™ 6,6 membrane made by Pall Biosupport, Port Washington, N.Y.)) or a polyvinylidine fluoride, such as Immobilon™ or Durapore™ membranes made by Millipore, Bedford, Mass. Other suitable membranes include, but are not limited to, cellulose, nitrocellulose, silica fiber, aluminum oxide, and polyvinyl chloride.

Binding elements may be immobilized on the selected membrane in any manner that assures the availability on the immobilized binding element of at least one binding site for selectively binding the labelled analyte and target analyte in an aqueous medium. Several methods for attaching binding elements to the membranes are well-known and therefore will not be specifically described herein. The binding element may be immobilized either throughout the thickness of the membrane, or on only one or both surfaces thereof.

The binding element may be any substance that can be immobilized on the membrane and that specifically binds the target analyte and its labelled analog. Binding elements include, but are not limited to, lectins, antibodies, antibiotics, and binding proteins other than antibodies and antibiotics.

Once the binding elements have been immobilized on the membrane, their available binding sites for selectively binding with the analyte will usually be essentially saturated with a labelled analog of the analyte (denoted herein as a “labelled analyte”). Saturation of the available binding sites with the labelled analyte enhances sensitivity by assuring that the maximum number of analyte molecules will displace labelled analytes, rather than binding directly to unoccupied binding sites.

The membrane may be oriented in any manner with respect to sample flow that allows the sample to flow past the complex of binding element and labelled analyte on the membrane over the desired interaction time. For example, the sample may flow through and essentially normal to the plane of the membrane. Alternatively, the membrane may be configured as a dipstick and the sample allowed to flow laterally through the membrane, for example by capillary action. In another alternative, the membrane support may be a hollow fiber configured so the sample flows along the hollow center before passing through the membrane. In any embodiment of the present invention, the flow of the sample through the membrane may be passive (i.e., gravitational or capillary flow) or active (flow resulting entirely or partly from the action of a flow pump, manual pressure, or vacuum).

Any label useful in assays for the analyte may be used to label the analyte. Fluorophores are particularly useful labels. Suitable fluorophores include, but are not limited to Fluorescein, Cadaverine, Texas Red™ (Molecular Probes, Eugene, OR) and Cyanine 5™ (BDS, Pennsylvania). If used, the fluorophore label is typically one that is detectable in the visible to near infrared range.

Once the sample has completed its interaction with the membrane having the immobilized binding element-labelled analyte thereon, the processed sample (e.g., the effluent from a sample column or the portion of the sample that has passed through and beyond the labelled portion of a test strip) is then analyzed to determine the concentration of displaced labelled analyte. The detection means for this analysis includes a readout for informing the user that a threshold amount of the label has been detected in the sample. When the label is fluorescent, the detection means also includes a light source for exciting the fluorophore-labelled analytes. The detection system can use various methods of optical measurement, including but not limited to a spectrophotometer, infrared spectrometer, fluorimeter, optical biosensor, or the eye.

The present invention is useful in the detection, in aqueous media, of any analyte that specifically binds to the binding element. The invention may be used, for example, to detect the presence of analytes in body fluids (blood, semen, saliva, urine, etc.), water, pharmaceutical preparations, environmental samples, aerosols, foods, and beverages. If the sample suspected of containing the analyte is originally in a viscous liquid, solid, gaseous state, the sample is preferably further dissolved in water before being exposed to the membrane.

Multiple binding elements for multiple analytes can be immobilized on a single membrane. Membranes containing the same or a different binding element can be arranged in stacks. Where multiple binding elements for multiple analytes are used, different labels on the labelled analytes can be used to distinguish which analyte is present.

FIG. 1 schematically shows a device 10 according to the present invention where the membrane is normal to sample flow. Membrane 12, with binding elements covalently bound or otherwise immobilized thereto and available binding sites saturated with a labelled analyte of the analyte, is positioned across column 14. An aqueous sample entering the top of column 14 flows through membrane 12. Analyte in the sample interacts with membrane 12 and displaces the labelled analyte from membrane 12. The labelled analyte, if it does not displace another labelled analyte or unlabelled analyte from the membrane, joins the effluent from column 14. The aqueous sample effluent from column 14 then enters line 16, which carries the effluent to detector 18 for detecting the presence of the labelled analyte in the effluent from column 14.

FIG. 2 shows an alternative embodiment of the present invention, where the membrane is also normal to sample flow. Porous membrane 102, with binding elements covalently bound or otherwise immobilized thereto and available binding sites saturated with a labelled analyte of the analyte, is positioned across column 104 having an open tip. To prevent the flow of sample between the outer edge of membrane 102 and the inner wall of column 104, the membrane typically extends fully across the width of column 104. The open tip of column 104 is inserted into the top of container 106 (typically through a septum (not shown)), which holds a sample suspected of containing the analyte. Suction means 105 can apply a vacuum to pull sample from container 106 through membrane 102 into column 104. Any label in the column may be detected by a detection means external to the column. To facilitate this external detection, column 104 is preferably transparent to, or includes a suitably placed window transparent to, the energy used for detection.

Although FIG. 2 shows the suction means as a plunger and column 104 as the syringe housing the plunger, other vacuum arrangements are possible. For example, FIG. 3 shows a design similar to that used by Vacuutainers™. Evacuated tube 204 has porous membrane 205, with binding elements covalently bound or otherwise immobilized thereto and available binding sites saturated with a labelled analyte of the analyte, thereacross. To prevent the flow of sample between the outer edge of porous membrane 205 and the inner wall of evacuated tube 204, the membrane typically extends fully across the width of evacuated tube 204. The open end of evacuated tube 204 is sealed by cap 206 having flange 208 extending about the rim of open end of tube 204. Tip 210 extends from cap 206 opposite to hollow needle 212, which also extends from cap 206. Needle 212 extends to near septum 214 when tube 204 is placed, with only slight pressure, within flange 208. Septum 214 maintains the vacuum in the portion 216 of tube 204. Although septum 214 is essentially impermeable to gas or liquid, it is punctured by needle 212 once tube 204 is fully inserted into flange 208. Upon the puncture of septum 214, the vacuum within portion 216 draws liquid from sample container 218 through tip 210, into hollow needle 212, through membrane 205 and into portion 216. Any label within portion 216 can be detected as with other embodiments of the invention. To assure that needle 212 does not puncture membrane 205, the distance between the bottom of septum 214 and the bottom of membrane 205 should be greater than the height of needle 212. This embodiment of the invention assures that the flow across membrane 205 is consistent from sample to sample.

Having described the invention, the following examples are given to illustrate specific applications of the invention including the best mode now known to perform the invention. These specific examples are not intended to limit the scope of the invention described in this application.

EXAMPLES Example 1 TNT Detection

To prepare the membranes, the monoclonal 11B3 antibody (mouse 1gG,) with specificity for TNT (trinitrotoluene) was immobilized onto the Immunodyne® ABC membrane with a pore size of 0.45 μm. The 11B3 antibody, 100 μl of a 2 nmol/ml solution in phosphate buffered saline (PBS), was attached to the membrane by either placing the solution in a test tube, with subsequent addition of the membrane, or pipetting the antibody into a column that already contained the membrane. Whether in a column or a test tube, membranes were incubated with the antibody for four hours at room temperature. Following incubation, the antibody solution was removed. Membranes exposed to antibody in a test tube were placed in a column. Any unreacted binding sites on the membrane were blocked with the addition of 100μl of 1M Tris for approximately 30 minutes. To reduce nonspecific binding, the membranes were drained and washed three times with PBS containing 0.01% Triton X-100® detergent.

The labelled analyte was prepared by attaching the fluorophore CY5® (BDS, Pennsylvania) to trinitrobenzyl cadaverine (CY5-TNB). To saturate the antibody binding site with the labelled antigen, a solution of the CY5-TNB (4 nmoles in 50 μl PBS) was added to each column, and the columns were placed on a rocking bed overnight. The columns were connected to the fluorimeter and, washed briefly. Samples were introduced at a flow rate of 1 mL/min. Analyte injections were made in triplicate with concentrations ranging between 18.75 ng/mL and 1200 ng/mL. FIG. 2 illustrates data obtained for a membrane assay prepared with the test tube incubation method. A fluorescence signal peak was obtained at all analyte concentrations which was proportional to the amount of analyte added to the column.

FIG. 3 represents data from a membrane assay prepared by saturating the immobilized antibody with labelled analyte in the column as opposed to in a test tube. Again, an increase in signal intensity with increasing analyte concentration was observed. However, a plateau was seen between an analyte concentration of 700 ng/mL and 1200 ng/mL where a negligible increase in signal intensity was observed despite a two-fold increase in analyte concentration suggesting that there is less labelled analyte on the membrane available for displacement, compared to the membrane prepared in the test tube.

Both FIGS. 3 and 4 demonstrate reproducible results, with minimal standard error as indicated by the error bars. Assay times were fast with the exact time being simply a function of the flow rate (1 mL/min in this case) and the length of tubing between the analyte introduction site and the fluorimeter flow cell. For these experiments, signals were generated less than 1 minute from the time of sample introduction.

Example 2 Detection of RDX

Similar experiments were conducted whereby a monoclonal antibody with specificity for the explosive, cyclonite (RDX), was immobilized onto the membrane. The procedure for immobilization was identical to the one used for the anti-TNT antibody. However, 100 μl of 0.5% casein was used instead of Tris in order to block the remaining binding sites on the membrane. FIG. 4 represents data from a single membrane assay prepared by saturating the antibody directly in the column. A linear relationship between signal intensity and analyte concentration is observed. The lower limit of detection for this assay is at 5 ng/ml which corresponds to part per billion (ppb) levels.

II. Displacement Dipstick Studies.

The main objective of these experiments was to design a qualitative membrane-based immunoassay for the detection of a target analyte in solution. The tests rely the displacement immunoassay to work on the Immunodyne membranes with the fluid flowing through them membranes laterally as opposed to perpendicular to the membrane as described above. Transported by capillary action, the fluid conducts the analyte in the sample to the immobilized antibody-labelled analyte complex and transports the displaced labelled analyte further along the membrane strip. The dipstick displacement assay is not only dependent upon the ability of the target analyte to displace the labelled analyte from the immobilized antibody but also on several other factors such as the rate of the capillary action of the mobile phase and the rates of transport of analyte and labelled analyte through the membrane.

FIG. 5 provides a schematic of the experimental protocol. First, in step (a), strips 100 were cut from an ABC Immunodyne® membrane 110 that were either 30×5 mm or 50×10 mm. A monoclonal antibody specific for TNT (11B3) in concentrations ranging from 2 to 10 nmol/ml was placed in 5 μL droplets onto the membrane strips and allowed to immobilize for thirty minutes. In step (b), strips 100 were then soaked, using test tube 112, in a Tris solution for about an hour to block any other covalent binding sites. A washing of membrane strips 100 followed that consisted of three consecutive exposures to PBS containing 0.01% Triton X-100 to wash away any excess TNT antibody (step (c)). After a final wash with PBS, CY5-TNB labelled analyte, in excess of five-to-thirty times the molar amount of antibody, was applied in 6.5 μL droplets onto the antibody and incubated overnight (step (d)). In step (e), strips 100 were then washed in PBS containing 2.5% ethanol, and 1% Tween 20™ for ten minutes in order to remove nonspecifically bound labelled analyte. In step (f), before drying, strips 100 were put into a solution of 100 mM trehalose dihydrate in phosphate buffer for ten minutes. Finally, in step (g) the strips were dried at room temperature. The displacement assay (step (h)) was conducted by dipping the end of membrane strip 100 in TNT solution 114, 116, 118, or 120 (of the concentration specified in FIG. 5, step (h)), and allowing capillary action to bring the target analyte up to the antibody/labelled analyte complex for displacement. A 650 nm laser (not shown) connected to a fluorescence detector was used to look for any displaced labelled analyte (Cy5-TNB) on membrane strip 100.

In the first experiment, TNT antibody at a concentration of 2 nmol/ml was placed at the center of a 3×0.5 cm, rectangular membrane strip and was saturated by five times excess CY5-TNB. The strip was then dipped into a sample solution containing 300 ng/ml TNT. FIG. 6a represents this strip when the dipped end is held under the laser first (left side). The higher plateau indicates the fluorescence from the CY5-TNB bound to the immobilized antibody. A shoulder is evident to the right of the higher plateau, indicating displacement of the labelled analyte from the antibody. FIG. 6b shows this same strip optically interrogated in the reverse direction where the dipped end is on the right. Another membrane strip, also having 2 nmol/ml of immobilized anti-TNT antibody was treated identically and exposed to the same 300 ng/ml TNT solution. After placing it under the laser with the dipped end on the right, the data shown in FIG. 6c was obtained. These experiments were conducted by manually moving the membrane strip along the laser path. “Time” on the x-axis refers to scanning time and has no relation to assay time.

Obviously, many modifications and variations of the present invention are possible in light of the above teachings. It is therefore to be understood that, within the scope of the appended claims, the invention may be practiced otherwise than as specifically described.

Claims (19)

What is claimed is:
1. A quantitative assay method for detecting a target analyte, comprising the steps of:
providing a porous membrane having binding elements immobilized thereon, each of said binding elements having at least one binding site capable of specifically binding to said target analyte;
exposing said binding sites to a labelled analog of the target analyte to form complexes of membrane-immobilized binding elements and labelled analogs;
pumping a first aqueous liquid sample, suspected of containing the target analyte, so as to flow said first liquid sample normal to and through said membrane having said complexes thereon, at a flow rate allowing the target analyte to displace the labelled analog from the complexes under non-equilibrium conditions to form downstream of said membrane a flowable liquid effluent including said displaced labelled analog, said flow rate also providing an interaction time between said analyte and said membrane of about 0.1 sec through about 30 sec;
iterrogating said flowable liquid effluent to detect and quantitatively determine the amount of the displaced labelled analog, the amount of said displaced labelled analog being proportional to the concentration of said target analyte in said first sample.
2. The method of claim 1, wherein the binding element is an antibody.
3. The method of claim 1, wherein said labelled analog is fluorescently labelled.
4. The method of claim 1, wherein said interaction time is no more than about 15 seconds.
5. The method of claim 1, wherein said membrane is nonabsorbent.
6. The method of claim 5, wherein said membrane is selected from the group consisting of cellulose, nitrocellulose, silica fiber, aluminum oxide, and polyvinyl chloride.
7. A device for the assay of an aqueous sample suspected of containing a target analyte, comprising:
a porous membrane having binding elements immobilized thereon, each of said binding elements having at least one binding site capable of specifically binding to said target analyte, essentially all of said binding sites on said membrane being occupied by a labelled analog of the target analyte to form complexes of membrane-immobilized binding elements and labelled analogs;
a pump for flowing an aqueous liquid sample, suspected of containing the target analyte, normal to and through said membrane having said complexes thereon, at a flow rate allowing the target analyte to displace the labelled analog from the complexes under non-equilibrium conditions to form downstream of said membrane a flowable liquid effluent including said displaced labelled analog, said flow rate also providing an interaction time between said analyte and said membrane of about 0.1 sec through about 30 sec; and
a detector that interrogates said flowable liquid effluent and the presence of said labelled analog therein.
8. The device of claim 7, wherein said labelled analog is fluorescently labelled.
9. The device of claim 8, wherein said detecting means further includes a light source for exciting any fluorescently labelled analog in said processed sample.
10. The device of claim 7, wherein said binding element is an antibody.
11. The device of claim 9, wherein said detecting means further is adapted to quantitatively determine the amount of said labelled analog in said processed sample.
12. The device of claim 9, wherein said detecting means further comprises a spectrophotometer, infrared spectrometer, fluorimeter or an optical biosensor.
13. The device of claim 7, wherein said membrane is nonabsorbent.
14. The device of claim 7, wherein said flow means is adapted to provide an interaction time between said analyte and said membrane of no more than about 15 sec.
15. The method of claim 1, further comprising, after said interrogating step, the steps of:
rinsing said sample from said membrane;
pumping a second aqueous liquid sample, suspected of containing target analyte, so as to flow said second liquid sample normal to and through said rinsed membrane having said complexes thereon, at a flow rate allowing said target analyte in said second sample to displace the labelled analog from the complexes under non-equilibrium conditions to form downstream of said membrane a flowable liquid effluent including said labelled analog displaced by said target analyte in said second sample, said flow rate also providing an interaction time between said target analyte in said second sample and said membrane of about 0.1 sec through about 30 sec;
interrogating said liquid effluent to detect and quantitatively determine the amount of the displaced labelled analog, the amount of said displaced labelled analog being proportional to the concentration of said target analyte in said second sample.
16. The device of claim 7, wherein, after said device has been used to assay a first aqueous sample suspected of containing said liquid analyte by:
flowing said first aqueous liquid sample, suspected of containing the target analyte, normal to and through said membrane having said complexes thereon, at a flow rate allowing the target analyte to displace the labelled analog from the complexes under non-equilibrium conditions to form downstream of said membrane a liquid effluent including said displaced labelled analog , said flow rate also providing an interaction time between said analyte and said membrane of about 0.1 sec through about 30 sec; and
interrogating said liquid effluent for the presence of said labelled analog therein,
said membrane may be rinsed and said device may be reused for the steps of:
flowing a second aqueous liquid sample, suspected of containing target analyte, normal to and through said membrane having said complexes thereon, at a flow rate allowing the target analyte in said second sample to displace the labelled analog from the complexes under non-equilibrium conditions to form downstream of said membrane a liquid effluent including said labelled analog displaced by said target analyte in said second sample, said flow rate also providing an interaction time between said analyte in said second sample and said membrane of about 0.1 sec through about 30 sec; and
interrogating said liquid effluent from said second sample for the presence of said labelled analog displaced by said target analyte in said second sample.
17. A continuous flow assay according to claim 1, wherein said sample is injected, upstream of said membrane, into a continuous stream of buffer flowing through said membrane.
18. A device according to claim 7, wherein said pump means causes a continuous stream of buffer to flow through said membrane, and further including a injector, upstream of said membrane, that injects said sample into said stream.
19. A continuous flow assay according to claim 15, wherein said first and second samples are injected, upstream of said membrane, into a continuous stream of buffer flowing through said membrane, wherein said rinsing step is performed by the action of said buffer stream between said step of pumping said first liquid sample and said step of pumping said second liquid sample.
US08/583,912 1996-01-11 1996-01-11 Displacement assay on a porous membrane Expired - Fee Related US6750031B1 (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
US08/583,912 US6750031B1 (en) 1996-01-11 1996-01-11 Displacement assay on a porous membrane

Applications Claiming Priority (8)

Application Number Priority Date Filing Date Title
US08/583,912 US6750031B1 (en) 1996-01-11 1996-01-11 Displacement assay on a porous membrane
PCT/US1996/016981 WO1997025619A1 (en) 1996-01-11 1996-10-18 Displacement assay on a porous membrane
AT96938636T AT327507T (en) 1996-01-11 1996-10-18 Displacement assay on a porous membrane
DE1996636173 DE69636173T2 (en) 1996-01-11 1996-10-18 Displacement assay on a porous membrane
EP19960938636 EP0880699B1 (en) 1996-01-11 1996-10-18 Displacement assay on a porous membrane
CA 2241324 CA2241324C (en) 1996-01-11 1996-10-18 Displacement assay on a porous membrane
DE1996636173 DE69636173D1 (en) 1996-01-11 1996-10-18 Displacement assay on a porous membrane
US09/907,888 US6808937B2 (en) 1996-01-11 2001-07-13 Displacement assay on a porous membrane

Related Child Applications (1)

Application Number Title Priority Date Filing Date
US09/907,888 Division US6808937B2 (en) 1996-01-11 2001-07-13 Displacement assay on a porous membrane

Publications (1)

Publication Number Publication Date
US6750031B1 true US6750031B1 (en) 2004-06-15

Family

ID=24335115

Family Applications (2)

Application Number Title Priority Date Filing Date
US08/583,912 Expired - Fee Related US6750031B1 (en) 1996-01-11 1996-01-11 Displacement assay on a porous membrane
US09/907,888 Expired - Fee Related US6808937B2 (en) 1996-01-11 2001-07-13 Displacement assay on a porous membrane

Family Applications After (1)

Application Number Title Priority Date Filing Date
US09/907,888 Expired - Fee Related US6808937B2 (en) 1996-01-11 2001-07-13 Displacement assay on a porous membrane

Country Status (5)

Country Link
US (2) US6750031B1 (en)
EP (1) EP0880699B1 (en)
AT (1) AT327507T (en)
DE (2) DE69636173T2 (en)
WO (1) WO1997025619A1 (en)

Cited By (19)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20040101900A1 (en) * 2002-11-25 2004-05-27 Mauro J. Mattew Assay for rapid detection of TNT
US20040241874A1 (en) * 2001-08-31 2004-12-02 Mohamed Abdel-Rehim Method and apparatus for sample preparation using solid phase microextraction
US20050126908A1 (en) * 2002-04-04 2005-06-16 Keenan Elizabeth A. Electrochemical detector systems
US20050158870A1 (en) * 2001-01-26 2005-07-21 Surromed, Inc. Surface-enhanced spectroscopy-active sandwich nanoparticles
US20050219509A1 (en) * 1999-10-06 2005-10-06 Natan Michael J Surface enhanced spectroscopy-active composite nanoparticles
US20060054506A1 (en) * 1999-10-06 2006-03-16 Natan Michael J Surface enhanced spectrometry-active composite nanoparticles
US20090155811A1 (en) * 2006-01-27 2009-06-18 Oxonica, Inc. Lateral Flow Immunoassay With Encapsulated Detection Modality
US20090298197A1 (en) * 2005-11-15 2009-12-03 Oxonica Materials Inc. Sers-based methods for detection of bioagents
US20100060893A1 (en) * 2006-07-24 2010-03-11 Norton Scott M Assay particle concentration and imaging apparatus and method
US7682801B2 (en) 2005-03-11 2010-03-23 Chembio Diagnostic Systems, Inc. Dual path immunoassay device
US7723100B2 (en) 2006-01-13 2010-05-25 Becton, Dickinson And Company Polymer coated SERS nanotag
US7879597B2 (en) 2005-03-11 2011-02-01 Chembio Diagnostic Systems, Inc. Dual path immunoassay device
US7879622B1 (en) * 2006-08-11 2011-02-01 University Of South Florida Barrier-permeable proxy reporter analysis
US7943395B2 (en) * 2003-11-21 2011-05-17 Kimberly-Clark Worldwide, Inc. Extension of the dynamic detection range of assay devices
US20110151479A1 (en) * 2008-08-25 2011-06-23 University Of Washington Microfluidic systems incorporating flow-through membranes
US8409863B2 (en) 2005-12-14 2013-04-02 Becton, Dickinson And Company Nanoparticulate chemical sensors using SERS
US8603835B2 (en) 2011-02-10 2013-12-10 Chembio Diagnostic Systems, Inc. Reduced step dual path immunoassay device and method
US9138743B2 (en) 2006-10-04 2015-09-22 University Of Washington Method and device for rapid parallel microfluidic molecular affinity assays
US9885710B2 (en) 2014-04-02 2018-02-06 Chembio Diagnostic Systems, Inc. Immunoassay utilizing trapping conjugate

Families Citing this family (82)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5891740A (en) * 1997-04-02 1999-04-06 The Perkin-Elmer Corporation Detection of low level hydrophobic analytes in environmental samples using agglutination reaction capillary slide test and apparatus therefor
US6036924A (en) 1997-12-04 2000-03-14 Hewlett-Packard Company Cassette of lancet cartridges for sampling blood
GB9726888D0 (en) * 1997-12-20 1998-02-18 Eev Ltd Detection
US6391005B1 (en) 1998-03-30 2002-05-21 Agilent Technologies, Inc. Apparatus and method for penetration with shaft having a sensor for sensing penetration depth
US8641644B2 (en) 2000-11-21 2014-02-04 Sanofi-Aventis Deutschland Gmbh Blood testing apparatus having a rotatable cartridge with multiple lancing elements and testing means
US7892183B2 (en) 2002-04-19 2011-02-22 Pelikan Technologies, Inc. Method and apparatus for body fluid sampling and analyte sensing
US8372016B2 (en) 2002-04-19 2013-02-12 Sanofi-Aventis Deutschland Gmbh Method and apparatus for body fluid sampling and analyte sensing
US8221334B2 (en) 2002-04-19 2012-07-17 Sanofi-Aventis Deutschland Gmbh Method and apparatus for penetrating tissue
US7331931B2 (en) 2002-04-19 2008-02-19 Pelikan Technologies, Inc. Method and apparatus for penetrating tissue
US7901362B2 (en) 2002-04-19 2011-03-08 Pelikan Technologies, Inc. Method and apparatus for penetrating tissue
US8579831B2 (en) 2002-04-19 2013-11-12 Sanofi-Aventis Deutschland Gmbh Method and apparatus for penetrating tissue
WO2002100251A2 (en) 2001-06-12 2002-12-19 Pelikan Technologies, Inc. Self optimizing lancing device with adaptation means to temporal variations in cutaneous properties
US7297122B2 (en) 2002-04-19 2007-11-20 Pelikan Technologies, Inc. Method and apparatus for penetrating tissue
US7749174B2 (en) 2001-06-12 2010-07-06 Pelikan Technologies, Inc. Method and apparatus for lancet launching device intergrated onto a blood-sampling cartridge
US8337419B2 (en) 2002-04-19 2012-12-25 Sanofi-Aventis Deutschland Gmbh Tissue penetration device
US9427532B2 (en) 2001-06-12 2016-08-30 Sanofi-Aventis Deutschland Gmbh Tissue penetration device
US9226699B2 (en) 2002-04-19 2016-01-05 Sanofi-Aventis Deutschland Gmbh Body fluid sampling module with a continuous compression tissue interface surface
US8267870B2 (en) 2002-04-19 2012-09-18 Sanofi-Aventis Deutschland Gmbh Method and apparatus for body fluid sampling with hybrid actuation
ES2352998T3 (en) 2001-06-12 2011-02-24 Pelikan Technologies Inc. Lanceta electric actuator.
US7976476B2 (en) 2002-04-19 2011-07-12 Pelikan Technologies, Inc. Device and method for variable speed lancet
US7229458B2 (en) 2002-04-19 2007-06-12 Pelikan Technologies, Inc. Method and apparatus for penetrating tissue
US7717863B2 (en) 2002-04-19 2010-05-18 Pelikan Technologies, Inc. Method and apparatus for penetrating tissue
US7226461B2 (en) 2002-04-19 2007-06-05 Pelikan Technologies, Inc. Method and apparatus for a multi-use body fluid sampling device with sterility barrier release
DE60234597D1 (en) 2001-06-12 2010-01-14 Pelikan Technologies Inc Device and method for taking blood samples
US9248267B2 (en) 2002-04-19 2016-02-02 Sanofi-Aventis Deustchland Gmbh Tissue penetration device
US9795334B2 (en) 2002-04-19 2017-10-24 Sanofi-Aventis Deutschland Gmbh Method and apparatus for penetrating tissue
US8360992B2 (en) 2002-04-19 2013-01-29 Sanofi-Aventis Deutschland Gmbh Method and apparatus for penetrating tissue
US7648468B2 (en) 2002-04-19 2010-01-19 Pelikon Technologies, Inc. Method and apparatus for penetrating tissue
US7232451B2 (en) 2002-04-19 2007-06-19 Pelikan Technologies, Inc. Method and apparatus for penetrating tissue
US9314194B2 (en) 2002-04-19 2016-04-19 Sanofi-Aventis Deutschland Gmbh Tissue penetration device
US7547287B2 (en) 2002-04-19 2009-06-16 Pelikan Technologies, Inc. Method and apparatus for penetrating tissue
US7041068B2 (en) 2001-06-12 2006-05-09 Pelikan Technologies, Inc. Sampling module device and method
US7674232B2 (en) 2002-04-19 2010-03-09 Pelikan Technologies, Inc. Method and apparatus for penetrating tissue
US7909778B2 (en) 2002-04-19 2011-03-22 Pelikan Technologies, Inc. Method and apparatus for penetrating tissue
US7491178B2 (en) 2002-04-19 2009-02-17 Pelikan Technologies, Inc. Method and apparatus for penetrating tissue
US7291117B2 (en) 2002-04-19 2007-11-06 Pelikan Technologies, Inc. Method and apparatus for penetrating tissue
US7981056B2 (en) 2002-04-19 2011-07-19 Pelikan Technologies, Inc. Methods and apparatus for lancet actuation
US8784335B2 (en) 2002-04-19 2014-07-22 Sanofi-Aventis Deutschland Gmbh Body fluid sampling device with a capacitive sensor
US7344507B2 (en) 2002-04-19 2008-03-18 Pelikan Technologies, Inc. Method and apparatus for lancet actuation
US7371247B2 (en) 2002-04-19 2008-05-13 Pelikan Technologies, Inc Method and apparatus for penetrating tissue
AT497731T (en) 2001-06-12 2011-02-15 Pelikan Technologies Inc Apparatus for increasing the success rate in respect of blood exploitation obtained by a fingerstick
US7232689B2 (en) * 2002-03-11 2007-06-19 Pawliszyn Janusz B Calibration procedure for investigating biological systems
JP4381820B2 (en) * 2002-03-11 2009-12-09 ポーリスツィン、ジャヌス・ビー Microextraction device for biopsy
US9733234B2 (en) 2002-03-11 2017-08-15 Jp Scientific Limited Probe for extraction of molecules of interest from a sample
US9870907B2 (en) 2002-03-11 2018-01-16 Jp Scientific Limited Probe for extraction of molecules of interest from a sample
US8598325B2 (en) 2002-03-11 2013-12-03 Janusz B. Pawliszyn Solid-phase microextraction coatings and methods for their preparation
US20090026122A1 (en) * 2002-03-11 2009-01-29 Janusz Biocompatible solid-phase microextraction coatings and methods for their preparation
US8702624B2 (en) 2006-09-29 2014-04-22 Sanofi-Aventis Deutschland Gmbh Analyte measurement device with a single shot actuator
JP2005528612A (en) * 2002-06-03 2005-09-22 パムジーン ビー.ブイ. A new method for monitoring biomolecular interactions
US8574895B2 (en) 2002-12-30 2013-11-05 Sanofi-Aventis Deutschland Gmbh Method and apparatus using optical techniques to measure analyte levels
US20040147619A1 (en) * 2003-01-23 2004-07-29 Conocophillips Company Chlorine-containing synthesis gas catalyst
ES2347248T3 (en) 2003-05-30 2010-10-27 Pelikan Technologies Inc. Method and apparatus for injecting fluid.
EP1633235B1 (en) 2003-06-06 2014-05-21 Sanofi-Aventis Deutschland GmbH Apparatus for body fluid sampling and analyte sensing
EP1671096A4 (en) 2003-09-29 2009-09-16 Pelikan Technologies Inc Method and apparatus for an improved sample capture device
WO2005037095A1 (en) 2003-10-14 2005-04-28 Pelikan Technologies, Inc. Method and apparatus for a variable user interface
EP1706026B1 (en) 2003-12-31 2017-03-01 Sanofi-Aventis Deutschland GmbH Method and apparatus for improving fluidic flow and sample capture
JP2007534936A (en) * 2004-02-11 2007-11-29 パムジーン ベー.フェー. Apparatus for analyzing an interaction between target and probe molecules
US20060257991A1 (en) * 2004-02-27 2006-11-16 Mcdevitt John T Integration of fluids and reagents into self-contained cartridges containing particle-based sensor elements and membrane-based sensor elements
US7781226B2 (en) * 2004-02-27 2010-08-24 The Board Of Regents Of The University Of Texas System Particle on membrane assay system
US20060257941A1 (en) * 2004-02-27 2006-11-16 Mcdevitt John T Integration of fluids and reagents into self-contained cartridges containing particle and membrane sensor elements
WO2005083423A2 (en) 2004-02-27 2005-09-09 Board Of Regents, The University Of Texas System System and method for integrating fluids and reagents in self-contained cartridges containing particle and membrane sensor elements
US8101431B2 (en) 2004-02-27 2012-01-24 Board Of Regents, The University Of Texas System Integration of fluids and reagents into self-contained cartridges containing sensor elements and reagent delivery systems
US8105849B2 (en) 2004-02-27 2012-01-31 Board Of Regents, The University Of Texas System Integration of fluids and reagents into self-contained cartridges containing sensor elements
EP1751546A2 (en) 2004-05-20 2007-02-14 Albatros Technologies GmbH & Co. KG Printable hydrogel for biosensors
US9820684B2 (en) 2004-06-03 2017-11-21 Sanofi-Aventis Deutschland Gmbh Method and apparatus for a fluid sampling device
US9775553B2 (en) 2004-06-03 2017-10-03 Sanofi-Aventis Deutschland Gmbh Method and apparatus for a fluid sampling device
WO2006001797A1 (en) 2004-06-14 2006-01-05 Pelikan Technologies, Inc. Low pain penetrating
US8652831B2 (en) 2004-12-30 2014-02-18 Sanofi-Aventis Deutschland Gmbh Method and apparatus for analyte measurement test time
US7822454B1 (en) 2005-01-03 2010-10-26 Pelikan Technologies, Inc. Fluid sampling device with improved analyte detecting member configuration
CA2610793A1 (en) 2005-05-31 2007-05-10 Labnow, Inc. Methods and compositions related to determination and use of white blood cell counts
US8409411B2 (en) * 2005-12-02 2013-04-02 State Of Oregon Acting By And Through The State Board Of Higher Education On Behalf Of Portland State University Nano-porous membrane based sensors
US20090047673A1 (en) * 2006-08-22 2009-02-19 Cary Robert B Miniaturized lateral flow device for rapid and sensitive detection of proteins or nucleic acids
US8613214B2 (en) * 2008-01-09 2013-12-24 Orono Spectral Solutions, Inc. Apparatus and method for determining analyte content in a fluid
US9386944B2 (en) 2008-04-11 2016-07-12 Sanofi-Aventis Deutschland Gmbh Method and apparatus for analyte detecting device
WO2009137059A1 (en) 2008-05-05 2009-11-12 Los Alamos National Security, Llc Highly simplified lateral flow-based nucleic acid sample preparation and passive fluid flow control
US9375169B2 (en) 2009-01-30 2016-06-28 Sanofi-Aventis Deutschland Gmbh Cam drive for managing disposable penetrating member actions with a single motor and motor and control system
US8965476B2 (en) 2010-04-16 2015-02-24 Sanofi-Aventis Deutschland Gmbh Tissue penetration device
US9795747B2 (en) 2010-06-02 2017-10-24 Sanofi-Aventis Deutschland Gmbh Methods and apparatus for lancet actuation
GB201017447D0 (en) 2010-10-15 2010-12-01 Moorlodge Biotech Ventures Ltd Assay device
EP2699698B1 (en) 2011-04-20 2017-01-04 Mesa Biotech, Inc. Oscillating amplification reaction for nucleic acids
GB201510850D0 (en) * 2015-06-19 2015-08-05 Cambridge Molecular Diagnositcs Ltd Nucleic acid amplification and detection assays
CA3019256A1 (en) 2016-05-10 2017-11-16 Jp Scientific Limited System and method for desorbing and detecting an analyte sorbed on a solid phase microextraction device

Citations (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4258001A (en) 1978-12-27 1981-03-24 Eastman Kodak Company Element, structure and method for the analysis or transport of liquids
US4859583A (en) 1985-02-25 1989-08-22 Amoco Corporation Chemiluminescent immunochemical technique for low molecular weight antigens
US4895809A (en) * 1984-01-09 1990-01-23 Varian Associates, Inc. Immobilized antigen-antibody displacement process
US4916056A (en) 1986-02-18 1990-04-10 Abbott Laboratories Solid-phase analytical device and method for using same
US4920046A (en) 1987-02-20 1990-04-24 Becton, Dickinson And Company Process, test device, and test kit for a rapid assay having a visible readout
US5045479A (en) * 1988-07-05 1991-09-03 The Johns Hopkins University Continuous flow competitive assay with reference system
US5183740A (en) 1990-02-23 1993-02-02 The United States Of America As Represented By The Secretary Of The Navy Flow immunosensor method and apparatus
US5206177A (en) * 1988-01-21 1993-04-27 Boehringer Mannheim Corporation Apparatus for determining an analyte and method therefor
US5340748A (en) * 1990-07-18 1994-08-23 Abbott Laboratories Analyte-substitute reagent for use in specific binding assay methods, devices and kits
US5354654A (en) * 1993-07-16 1994-10-11 The United States Of America As Represented By The Secretary Of The Navy Lyophilized ligand-receptor complexes for assays and sensors
US5369007A (en) 1990-09-07 1994-11-29 The United States Of America As Represented By The Secretary Of The Navy Microassay on a card
US5470713A (en) * 1989-04-26 1995-11-28 Diagnostic Products Corporation Method and element for measuring analytes in biological fluids using immobilized binder-analyte labeled complex
US5573921A (en) * 1992-09-04 1996-11-12 Dragerwerk Aktiengesellschaft Immunochemical displacement for determining an analyte

Family Cites Families (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3800780A (en) * 1972-02-23 1974-04-02 Angelika Elliott Vacuum indicator
US4812293A (en) * 1986-06-30 1989-03-14 Becton, Dickinson And Company Vacuum actuated assay device and method of using same
US5024238A (en) * 1989-01-10 1991-06-18 Cancer Diagnostics, Inc. Blood withdrawing apparatus and antigen testing method
US5602037A (en) * 1994-06-30 1997-02-11 Dade International, Inc. Combination reagent holding and test device
US6016712A (en) * 1997-09-18 2000-01-25 Accumetrics Device for receiving and processing a sample

Patent Citations (13)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4258001A (en) 1978-12-27 1981-03-24 Eastman Kodak Company Element, structure and method for the analysis or transport of liquids
US4895809A (en) * 1984-01-09 1990-01-23 Varian Associates, Inc. Immobilized antigen-antibody displacement process
US4859583A (en) 1985-02-25 1989-08-22 Amoco Corporation Chemiluminescent immunochemical technique for low molecular weight antigens
US4916056A (en) 1986-02-18 1990-04-10 Abbott Laboratories Solid-phase analytical device and method for using same
US4920046A (en) 1987-02-20 1990-04-24 Becton, Dickinson And Company Process, test device, and test kit for a rapid assay having a visible readout
US5206177A (en) * 1988-01-21 1993-04-27 Boehringer Mannheim Corporation Apparatus for determining an analyte and method therefor
US5045479A (en) * 1988-07-05 1991-09-03 The Johns Hopkins University Continuous flow competitive assay with reference system
US5470713A (en) * 1989-04-26 1995-11-28 Diagnostic Products Corporation Method and element for measuring analytes in biological fluids using immobilized binder-analyte labeled complex
US5183740A (en) 1990-02-23 1993-02-02 The United States Of America As Represented By The Secretary Of The Navy Flow immunosensor method and apparatus
US5340748A (en) * 1990-07-18 1994-08-23 Abbott Laboratories Analyte-substitute reagent for use in specific binding assay methods, devices and kits
US5369007A (en) 1990-09-07 1994-11-29 The United States Of America As Represented By The Secretary Of The Navy Microassay on a card
US5573921A (en) * 1992-09-04 1996-11-12 Dragerwerk Aktiengesellschaft Immunochemical displacement for determining an analyte
US5354654A (en) * 1993-07-16 1994-10-11 The United States Of America As Represented By The Secretary Of The Navy Lyophilized ligand-receptor complexes for assays and sensors

Non-Patent Citations (7)

* Cited by examiner, † Cited by third party
Title
"Enzyme", Analytical Chemistry, vol. 56, No. 8, , Jul. 1984, 920A-922A, 926A, 928A, 930A-931A.
Curme et al., Clin. Chem., 24/8, 1335-1341.
Granite Diagnostics, Inc, "Description of the E-Z Screen Test System" Brochure, Oct. 7, 1985.
Greener et al., Clin. Chem., 34, 1865 (1989).
Ramp (TM) Urine hcG Assay brochure (1986), RMP00230 M 0186.
Ramp ™ Urine hcG Assay brochure (1986), RMP00230 M 0186.
Wemhoff et al., J. Immunol. Methods, 156 (1992) 223-230.

Cited By (32)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20060054506A1 (en) * 1999-10-06 2006-03-16 Natan Michael J Surface enhanced spectrometry-active composite nanoparticles
US8497131B2 (en) 1999-10-06 2013-07-30 Becton, Dickinson And Company Surface enhanced spectroscopy-active composite nanoparticles comprising Raman-active reporter molecules
US20110172523A1 (en) * 1999-10-06 2011-07-14 Oxonica, Inc. Surface enhanced spectroscopy-active composite nanoparticles
US9201013B2 (en) 1999-10-06 2015-12-01 Becton, Dickinson And Company Method for tagging material with surface-enhanced spectroscopy (SES)-active composite nanoparticles
US20090121193A1 (en) * 1999-10-06 2009-05-14 Oxonica, Inc. Surface enhanced spectroscopy-active composite nanoparticles
US20050219509A1 (en) * 1999-10-06 2005-10-06 Natan Michael J Surface enhanced spectroscopy-active composite nanoparticles
US8918161B2 (en) 1999-10-06 2014-12-23 Becton, Dickinson And Company Methods of use for surface enhanced spectroscopy-active composite nanoparticles
US20050221494A1 (en) * 2001-01-26 2005-10-06 Natan Michael J Surface-enhanced spectroscopy-active sandwich nanoparticles
US20050208663A1 (en) * 2001-01-26 2005-09-22 Natan Michael J Surface-enhanced spectroscopy-active sandwich nanoparticles
US9297766B2 (en) 2001-01-26 2016-03-29 Becton, Dickinson And Company Method of tagging materials with surface-enhanced spectroscopy-active sandwich particles
US20050158870A1 (en) * 2001-01-26 2005-07-21 Surromed, Inc. Surface-enhanced spectroscopy-active sandwich nanoparticles
US20040241874A1 (en) * 2001-08-31 2004-12-02 Mohamed Abdel-Rehim Method and apparatus for sample preparation using solid phase microextraction
US7547384B2 (en) 2002-04-04 2009-06-16 Millipore Corporation Electrochemical detector systems
US20050126908A1 (en) * 2002-04-04 2005-06-16 Keenan Elizabeth A. Electrochemical detector systems
US20040101900A1 (en) * 2002-11-25 2004-05-27 Mauro J. Mattew Assay for rapid detection of TNT
US7943395B2 (en) * 2003-11-21 2011-05-17 Kimberly-Clark Worldwide, Inc. Extension of the dynamic detection range of assay devices
US7879597B2 (en) 2005-03-11 2011-02-01 Chembio Diagnostic Systems, Inc. Dual path immunoassay device
US8507259B2 (en) 2005-03-11 2013-08-13 Chembio Diagnostics Systems, Inc. Dual path immunoassay device
US9784734B2 (en) 2005-03-11 2017-10-10 Chembio Diagnostic Systems, Inc. Dual path immunoassay device
US7682801B2 (en) 2005-03-11 2010-03-23 Chembio Diagnostic Systems, Inc. Dual path immunoassay device
US8877450B2 (en) 2005-03-11 2014-11-04 Chembio Diagnostic Systems, Inc. Dual path immunoassay device
US20090298197A1 (en) * 2005-11-15 2009-12-03 Oxonica Materials Inc. Sers-based methods for detection of bioagents
US8409863B2 (en) 2005-12-14 2013-04-02 Becton, Dickinson And Company Nanoparticulate chemical sensors using SERS
US7723100B2 (en) 2006-01-13 2010-05-25 Becton, Dickinson And Company Polymer coated SERS nanotag
US20090155811A1 (en) * 2006-01-27 2009-06-18 Oxonica, Inc. Lateral Flow Immunoassay With Encapsulated Detection Modality
US20100060893A1 (en) * 2006-07-24 2010-03-11 Norton Scott M Assay particle concentration and imaging apparatus and method
US7879622B1 (en) * 2006-08-11 2011-02-01 University Of South Florida Barrier-permeable proxy reporter analysis
US9138743B2 (en) 2006-10-04 2015-09-22 University Of Washington Method and device for rapid parallel microfluidic molecular affinity assays
US20110151479A1 (en) * 2008-08-25 2011-06-23 University Of Washington Microfluidic systems incorporating flow-through membranes
US8603835B2 (en) 2011-02-10 2013-12-10 Chembio Diagnostic Systems, Inc. Reduced step dual path immunoassay device and method
US9885710B2 (en) 2014-04-02 2018-02-06 Chembio Diagnostic Systems, Inc. Immunoassay utilizing trapping conjugate
US9891216B2 (en) 2014-04-02 2018-02-13 Chembio Diagnostic Systems, Inc. Immunoassay methods utilizing trapping conjugate

Also Published As

Publication number Publication date
EP0880699A4 (en) 2002-04-10
AT327507T (en) 2006-06-15
EP0880699A1 (en) 1998-12-02
DE69636173T2 (en) 2007-03-29
DE69636173D1 (en) 2006-06-29
US20020028475A1 (en) 2002-03-07
EP0880699B1 (en) 2006-05-24
WO1997025619A1 (en) 1997-07-17
US6808937B2 (en) 2004-10-26

Similar Documents

Publication Publication Date Title
US5569608A (en) Quantitative detection of analytes on immunochromatographic strips
EP0191640B2 (en) Concentrating immunochemical test strip
US10415104B2 (en) Devices for the detection of multiple analytes in a sample
US4496654A (en) Detection of HCG with solid phase support having avidin coating
EP0281201B1 (en) Self contained immunoassay element
EP0306336B2 (en) Multiple port assay device
US5418142A (en) Glucose test strip for whole blood
US6316205B1 (en) Assay devices and methods of analyte detection
KR100635110B1 (en) Lab-on-a-chip for an on-the-spot analysis and signal detector for the same
US7935308B2 (en) Methods and devices for analyte detection
EP0032286B1 (en) Method for analysis for a member of an immunological pair using a test surface; kit and test surface material therefor
JP3543000B2 (en) Biosensor
US5079142A (en) Orthogonal flow immunoassays and devices
US6436722B1 (en) Device and method for integrated diagnostics with multiple independent flow paths
US5281539A (en) Immunoassay device for continuous monitoring
US6245296B1 (en) Flow immunosensor apparatus
CA1272127A (en) Solid phase system for use in ligand-receptor assays
US5234813A (en) Method and device for metering of fluid samples and detection of analytes therein
US6991912B2 (en) Systems and methods for performing magnetic chromatography assays
ES2449491T3 (en) Method to immobilize conjugates in diagnostic tests
US5541069A (en) Assay having improved dose response curve
EP0319294B1 (en) Improved membrane assay using focused sample application
KR101262407B1 (en) Metering technique for lateral flow assay devices
US7632687B2 (en) Hybrid phase lateral flow assay
US20020019062A1 (en) Assay devices

Legal Events

Date Code Title Description
AS Assignment

Owner name: NAVY, THE UNITED STATES OF AMERICA AS REPRESENTED

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:LIGLER, FRANCES L.;KUSTERBECK, ANNE W.;RABBANY, SINA Y.;REEL/FRAME:007901/0293;SIGNING DATES FROM 19960126 TO 19960129

FPAY Fee payment

Year of fee payment: 4

REMI Maintenance fee reminder mailed
LAPS Lapse for failure to pay maintenance fees
STCH Information on status: patent discontinuation

Free format text: PATENT EXPIRED DUE TO NONPAYMENT OF MAINTENANCE FEES UNDER 37 CFR 1.362

FP Expired due to failure to pay maintenance fee

Effective date: 20120615